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Lecture Seamus Holden


17 November 2010 FWN-Building 5116.0116, Nijenborgh 4, 9747 AG, Groningen
Speaker: Dr. Seamus Holden
Affiliation: University of Oxford, United Kingdom
Title: Close to the edge: single-molecule FRET and imaging at the limits of resolution
Date: Wed Nov 17, 2010
Start: 11.00
Location: FWN-Building 5116.0116
Host: A. van Oijen
Telephone: +31 50 363 9883


The study of molecular machines such as the E. coli repair polymerase, DNA polymerase I (DNAP), is inherently a multiscale problem. On the nanoscale, we wish to understand DNAP as a single machine, characterised by distance fluctuations on the scale of one DNA base pair (0.34 nm). On the microscale we are interested in the spatial organisation of multiple DNAPs within the cell. This requires cutting-edge resolution in both regimes.

On the nanoscale, we extended the limits to resolution of smFRET carried out by total internal reflection fluorescence (TIRF) microscopy to the level of one DNA base pair, and characterised the major limiting factors to ultimate resolution.

On the microscale, we present a new approach (adapted from astronomy), which dramatically improves the performance of the algorithms used to analyse super-resolution microscopy data. Our approach improves accuracy, reduces image acquisition times and increases the ultimate spatial resolution of the technique. We are currently working towards application of these two methodological paradigms: to study DNAP translocation by smFRET, and intra-cellular DNAP spatial organisation by super-resolution imaging.

Last modified:22 October 2012 2.30 p.m.